Process for the manufacture of beta-carotene



(1) Synthesis of quadrupZy-unsaturated Cia-aldehyde The condensation of 4- [2,6,6trimethylcyclo hexene (1') yl] 2 methyl butene (2) -al-(l) with the 1-ethers of 1-hydroxy-2- methyl-butene-(l) -yne- (3) or 1,2-dihydroxy-2- methyl-butyne-(S) can be carried out by an alkali metal condensation as well as by a Grignard reaction. The condensation over the lithium compounds in ether solution is particularly suitable. If a hydroxyl group is present, the same is rst protected by lithium. The hydrolysis oi the .salts of the condensation product formed can be carried out by means of a saturated ammonium chloride solution.

The transformation of the resulting condensation product of the formula CH3 CH3 CH2-CH=cli-CH-CECX0R li E CH3 in which X means one of the groups CH3 --CHT- te ::on-

and R represents a hydrocarbon radical, into the Cis-aldehyde compounds can be effected by allyl rearrangement, dehydration and acid treatment, whereby 1 mol of hydrogen may be added onto the triple bond after the allyl rearrangement. For such purpose 1methoxy2,6dimethyl-2,5- dihydroxy 8 [2,6,6 trimethylcyclohexene (1')-yl]octene 6)yne(3) may be rearranged by allowing the compound to stand for about 48 hours in alcoholic solution with 1 per cent. aqueous sulphuric acid to produce l-methoxy- 2,6 dimethyl 2,7 dihydroxy 8 [2,6,6 trimethylcyclohexene (1') yl] octene (5) yne-(3) the addition of 1 mol of hydrogen onto the triple bond by catalytic hydrogenation in methanol solution with a little active palladium lcatalyst yields 1-methoxy-Z,6-dimethyl-2,7-dihydroxy 8 [2,6,6 trimethylcyclohexene '(1') -yll -octadiene-(3,5) by dehydration, for instance by boiling with p-toluene sulphonic acid in toluene solution or with phosphorous oxyvchloride in toluene-pyridine solution, the lmethoxy 2,6 dimethyl 8 i2,6,6 trimethylcyclohexene (1') yl] octatetraene (1,357) is formed and the acid cleavage of this enol ether leads, under rearrangement, to 2,6- dimethyl 8 -v [2,6,6' trimethylcyclohexene v(1') -yl] octatriene-(2,4,6) -al-(l) The same aldehyde may also be obtained in a similar manner from 1 methoxy 2,6 dimethyl 5 hydroxy 8 [2,6,6 trimethylcyclohexene (1') yl] octadiene-(l,6) yne-(3).

2,6 dimethyl 8 [2,6',6' trimethylcyclo hexene (1') yl] octadiene (5,7) yne (3)-al-(1) is formed from 1-methoxy2,6di methyl 2,5 dihydroxy 8 [2',6',6 trimethylcyclohexene (1') yl] octene (6) yne-(3) by allyl rearrangement and dehydration by means of phosphorous oxychloride and pyridine to toluene solution and acid treatment of the 1-methoXy-2,6dimethyl 8 [2',6,6' trimethylcyclohexene- (1') -yll octatriene-(1,5,'l) yne-(3). The same aldehyde can also be obtained from 1-metl1oxy-2,6dimethyl5-hydroxy- 8 [2',6',6trimethylcyclohexene (1') yl] octadiene -(1,6)yne(3) by allyl rearrangement, dehydration and subsequent acid treatment.

(2) Preparation of the dihydroy compound with 40 carbon atoms 1 mol of acetylene dirnagnesium bromide or lithium acetylide is reacted with 2 equivalents of 2,6 dimethyl 8 [2',6',6' trimethylcyclohexene (1') y1] oetatriene (2,45) al (1). The condensation can be eiected in the usual solvents, or instance, ethyl ether. The magnesiumor lithium salt formed is hydrolyzed in the usual way, for instance, with an ammonium chloride solution or with dilute sulphuric acid. The 1,18-di- [2 ,6 ,6 -trimethylcyclohexene (1') yl] 3,7,12,16tetramethyl 8,11 dihydroxy octadecahexaene (2,4,6,12,14,16) yne (9) (hereinafter called dihydroxyhexenine compound) obtained can be puried by means of liquid-liquid extraction or by chromatography. In the chromatogram it shows approximately the same adhesiveness as Xanthophyll; it cannot be extracted from 92 per cent. methyl alcohol by means of petroleum ether. The pure compound possesses a molecular weight of 570. As evidenced by complete hydrogenation and determination according to Zerewitinofi, the said compound contain 8 double bonds, 1 triple bond and 2 hydroxyl groups.

ln precisely the same manner, the preparation and the purification of the 1,18di[2',6',6'tr1 methylcyclohexene (1') 'yl] 3,7,12,16 tetramethyl 8,11 dihydroxy octadecatetraene (l,3,15,17)triyne(5,9,13) (hereinafter called dihydroxytetraenetriyne compound) from 2 mols of 2,6-dimethyl-8-[2',6,6'-trimethylcyclohexene- (1') yl] octadiene'- (5,7) yne (3) al- (l) and acetylene dimagnesium bromide or lithium acetylide may be eected.

(3) Transformation to ,I3-carotene The above named dihydroxyhexenine compound with 40 carbon atoms is dehydrated with a View to introducing two new double bonds. The dehydration is suitably effected with p-toluene sulphonic acid 1n boiling toluene. This dehydration process is preceded by allyl rearrangement. The hydrocarbon compound formed (9,9dehy dro--carotene, i. e. L18-di-[2,6',6'trimethyl cyclohexene (1') yl] 3,7,12,16 tetramethyloctadecaoctaene-(1,3,5,7,11,13,15,17) yne (9)) can be extracted from 92 per cent. methyl alcohol with petroleum ether. The unreacted dihydroxyhexenine compound which remains behind in the methyl alcoholic solution, can be isolated and once more subjected to dehydration.

The 9,9dehydrocarotene obtained by splitting ofi water is, suitably after purification, subjected to partial hydrogenation at the triple bond. This may be achieved in the same way as eiective for other polyenes, for instance, by catalytic hydrogenation in the presence of palladium catalysts, whereby the activity of the palladium has been reduced by the addition of lead and quinoline. The introduction of hydrogen is interrupted after 1 mol of hydrogen has been taken up. The ,tl-carotene thus obtained is isomerized, if necessary, by treatment with iodine, heat or light. By working up carefully the hydrogenation product, 9,9'rnonociscarotene is first obtained in a very'good yield,l which is' i'somerized advantageouslyf by Warming or, preferably by exposure to light, tov produce all-trans-Q-carotene.

The partial hydrogenati'on off thetriple bond can also be carried out prior todehydration. The transformation to -carotenethen follows the following'course (withsma-ller-yield) :y Partial hydrogenation of the triple bond,A dehydration and, in given cases, isomeri'zation. The methods for the execution' ofthe-single steps are substantially the same asdescribed above.

In an analogous manner, the dihydroxytetraenetriyne compound with40-carbcn atoms can-be transformed toy /S-carotene. The measures tobe taken in the singlel steps are thef'same asdescribed for the dihydroxyhexenine compound. No allyl rearrangement takes placeY during the dehydrationA and during the partial hydrogenation the introduction of hydrogen is interrupted after 3 mole of' hydrogenhave been taken up. The yields` of this modificationY are considerably smaller than those attained by thezpreferred: way of' synthesis. Y

According to the-present invention as setforth above synthetic ,o-caroteneis obtained which can be puried by thesame` methodsas employed for carotene concentrates prepared from: natural sources. (Separation byliquid-liquid extraction, chromatography and crystallization.) It is recommcndableto add antioxydants whichv may also be present' during theV whole course of the` synthesis; tocopherols` are especially suitable for this purpose.' In working up the products of the synthesis, cis-forms of the ecarotene can be-isolated which, by treatment with iodine, heat or exposure-tolight, can beisomerized toohtain Ialltrans-carotene. The isomerization by the aid of iodine, which leads to=a mixture` oi?v isomers, is known. The isomerization by' exposure to light takes a uniform course in thecase of the 9,9- cis--carotene; it is suitably carried` out in` an inert solvent, such asv hexane. The rearrangement isconsiderably quicker with day-light than with the light of a mercury'lamp. ment by` heating and exclusion of light may be effected with the dissolved" as well as with the melted substance; in the case of the 9,9-ci`scarotene, an equilibriumlis-already attained after 11/2. hours at 70 C., at which all-trans--carotene is predominant.

The all-trans--carotene obtained` directly or by isomerization. corresponds in all itsproperties with -carotene obtained' from natural. materials and, in particular, it' possesses the same biological activity.

EXAMPLE 1 In a 2Y liter three-necked ilask provided with stirrer, dropping funnel and reflux condenser a solution of lithium phenyl inV 500- cc. of ether (content: 3.7 grams ofi lithium=-55 mois.) is added dropwise,v invav nitrogen; atmosphere,` vto 30.7 grams (0.27 mols) `of I-methoxy-Z-methyl- 2-hydroxy-butyne-(3) in'60 co. of absolute ether. The pasty reaction mixture is heated.- to boiling for 45 minutes and' after cooling 48v grams (0:23 mols)` of 4- [2,6,6trimethylcyclohexene-( 1')v yll2methylbutene(2),-al-(1). in 85 cc'.V of absolute ether are added. The white precipitate dissolves slowly. To terminatey the reaction, the solution, after standingv overnight, is heated to boiling. for 1/2' hour.V After cooling down,V the mixture is decomposed with a solution of 50 grams of ammonium chloride, whilecooling;V the ethersolution' is washedl with water 'and-fdried' The rearrangeyne-(S) are dissolved over sodium sulphate brown-red oil 'rei maining behind atten" the evaporation-of the ether is dissolvedl in 550 cc. offpetroleumffether and shakenout four`L times with f8@ caca-chof 80 percent. methanol. They united methanol extracts are once* again washed with petroleum ether and 2 liters or waterare added. The separated'oil is taken up` in-petroleum ether (boiling point '501C2) and theaqueous4 layer is sha-kenout twice wi -ha mixture of petroleum ether and`- ether (4:1). The-*united petroleum ethersolutions'V are washed;witlrT water, dried and evaporated. 60= grams (='76' per cent. ofthe theoretical quantity) of I-methoxy- 2,6-dmethyl'- 2,5 dihydroxy` 8 trimethylcyelohexenyl octene-(fD--yne-(B) are obtained Vasa red oil". The same is dissolvedv in 450" cci.f ofpetroleum ether andA stirred for` 24" hours under Anitrogen at -30" C; 23-.8` grams (32per cent of the theoretical quantity) of white crystals are then separated which can bel recrystallizedr from petroleum ether (boiling point 60-80 C1). (Since theV above mixture is a mixture of isomers, no sharp melting point can begiven.) l-'nV addition, grams ofa viscousoil, which does-not crystallize, are obtained from thepetroleumether solution. The determination according' to ZerewitinolifV shows 2A atoms of active-hydroge ultraviolet absorption: no absorption vabove 230- mp. For the further treatment' the crystallized and oily 1 methoxy -'2,6'- dimethyl 2,5- dihydroxy- 8' trimethylcyclohexenyl -voctene (6)v yne (3) are employed.

50' grams of I-rnethoxy-Z;6-diinethyl-2,5-dilfiyi droXy -8 trmetliylcyclclnaxenyl-octene (16)' I liter of alcoholand 250 cc; of 5 per cent. sulphuri'c acid are added. After 48 hours, the mixture-is pouredinto water, extracted with ether', washedl neutral, dried' and evaporated. The l-methoxy-Z,6-dimethyl-2-,7 dihydroxy-S- [2 ,6 ,6' -trimcthylcyclohexenel" yll-octene- (5) -yne- (3) remains behindas` an al'- most colourless viscous oil. '(TJltr-aviol'et maximum: \=233 ma (6:15500').

grams of 1-methoxy-Z-dimethyl-ZH-dihydroXy-S- [2 ,6' ,6' -trimetliylcyclohexene- (10G-yl]- octene(5)-yne-(3')' are-dissolved in 200 cc. of methanol and allowed testa-nd overnight with 20 grams of platinized' charcoal (f4 per cent.)` in order to remove catalyst" poisons. The4 coal' is sucked 01T and washed outvwithv 1150 cc; of methanol. Thereupon; 4 grams. of' quinoline are added and the mix-turei's hydrogenated with 4 grams of palladium charcoal (4 per cent.) until 1 mol of hydrogen has been taken up. After sucking off the catalyst,the hydrogenation prod?- uct is washed with ether, water is added andthe mixture is extracted with ether. The ether solution is washed neutral', dried! and boiled down'. The l-methoxy-2,6dimethyl-2,7-dihydroXy-8- ['2"',6,6" trimethyl' -A c-yclohexenc (I"`)- yll octadiene-(3,5) [a viscous` yellow oil; ultraviolet maximum: x=242 mp. ('e=:1l1000)=]` thus obtained is dissolved in 250 cc. of toluene, placed in afiiask equipped with ak descending condenser, heated to boiling, whereupon a solution of 400 mg. of toluene sulphonic acid' in'Y hot. toluene is added. The solution immediately boils more intensively and turns brown-red, while water distills oi azeotropically. The solution is' kept boiling for 15 minutes, coole-d, washed' neutral with bicarbonate solutionv and water successively-,- dried and concentrated in vacuo.

The concentrate, 1'methoxy-zf-dimethyls# tatetraene(1,3,5,'7) -is diluted Ywith petroleum ether for purification purposes vand filtered through 100 grams of aluminium oxide. The ltrate is boiled down in vacuo and to cleave the ether 20 times its quantity ofA alcohol and the same quantity of 20 per cent. sulphuric acid are added. After standing for 12 hours at room temperature, water is added to the reaction solution, whereupon the same is extracted with ether and the ether solution is washed neutral, boiled down and chromatographed. When eluting with petroleum ether, the aldehyde remains behind in the column. while side products will be found in the, filtrate. Finally, the column is eluted with petroleum ether with an addition of 1 per cent. acetone. Upon concentration, 24 grams of 2,6-dimethyl-8-[2,6,6-trimethylcyclohexene- (1') -yll -octatriene- (2,4,6) -al- (1) may thus be isolated. Ultraviolet-absorption maximum at 328 mp..

8 grams of 2,6dirnethyl8[2,6,6trimethyl cyclohexene (1') yl] octatriene (246) al- (1) are dissolved in 20 cc. of ether and l5 cc. of acetylene-dimagnesium bromide are added to the solution. The mixture is shaken for 12 hours in a brown flask.v Thereupon, ammonium chloride solution is added, the ether solution is washed, dried and boiled down. The residue is taken up in petroleum ether and chromatographed at an inactivated aluminum oxide column. Dark-coloured side products may be eluted with petroleum ether-acetone (201). The L18-di- [2,',6 trimethylcyclohexene (1') yll 3,7,12,16 tetramethyl 8,11 dihydroxy octadecahexaene- (2,4,6,12,14,161) -yne- (9) is eluted with ether with'addition of 5 per cent. glacial acetic acid, and the eluate is washed neutral with Water, dried and boiled down.

For dehydration purposes, the residue is dissolved in 35 cc. of toluene and heated to boiling for 15 minutes with a solution of 50 mg. of toluene sulphonic acid in l cc. of toluene. The dark red solution t is thereafter diluted with petroleum ether, washed neutral and boiled down in vacuo. The residue is taken up in petroleum ether and chromatographed at an aluminium oxide column (activity grade I-1I). It is developed with petroleum ether-benzene (10:1) and a yellow rst running is separated as eluate. Thereupon dirty-coloured parts are removed from the upper end of the column, and the broad orange-red zone is eluted with absolute ether. After evaporation in vacuo, the veluate leaves behind a dark red oil which is dissolved in carbon disulphide. vAlcohol is then added and the solution concentrated in vacuo until it begins to become cloudy. After standing for l2 hours at 10 C. the 9,9p-dehydro--carotene crystalliaes out. Upon recrystallization from benzene-methanol, the red needles melt at 147 C. (in the evacuated tube). Ultraviolet-absorption maximum: )(:436 my. (6:105'000).

For purification purposes 113 mg. o 9,9de hydro--carotene are dissolved in l0 cc. of benzene (free from thiophene) and shaken with 200 mg. of Pt-SiOz (13 per cent. platinum on silica gel). The catalyst is sucked ofi and washed three times with l0 cc. each of benzene. The united solutions are concentrated under reduced pressure and, for the purpose of hydrogenation, benzene is added to obtain a volume of 10 ec. Upon addition of 0.05 gram of quinoline, the mixture is hydrogenated in the presence of 300 mg. of Pd-CaCOs-catalyst inactivated with lead, until 5 cc. of Hz have been taken kup (calculated for 1 mol H2=5.1 cc.). The catalyst is filtered off and washed out with petroleum ether. The red solutions are washed with 5 per cent. Sulphuric acid and water and concentrated to 5 cc. In order to isomerize the cis-forms of the lf3-carotene, the concentrate is dissolved in cc. of petroleum ether of boiling point 40-60 C., 2 mg. of iodine in 5 cc. of benzene are added and the mixture is heated to boiling for 30 minutes. Thereupon the mixture is concentrated under reduced pressure to 5 cc., 20 cc. of petroleum ether are added and the mixture is chromatographed at an aluminium oxide column (4 x 35 cm, activity grade IV). It is eluted with petroleum ether of boiling point 40-60 C. The rst fraction contains 9,9dehy dro-,B-carotene. Crystals of melting point 170 C. can be isolated from the second fraction. The

third fraction yields dark violet-red crystals of vmelting point 172 C. which, upon recrystallization from carbon disulphide-alcohol, melt at ITS-179 C. in the evacuated tube. They consist of carotene. The melting point o the mixture with natural -carotene (melting point 179 C.) does not shown any depression; the chloroform solution shows with antimonytrichloride a blue coloration which is typical for -carotene and the ultraviolet spectrum is of the picture as characteristic of -carotene.

EXAMPLE 2 Z3 grams of 2,6-dimethyl-8-[2,6',6'trimethyl cyclohexene (1') yll octatriene (2,4,6) al- (l), prepared according to Example 1, are dissolved in 50 cc. of absolute ether and shaken with 30 cc. of acetylene dimagnesium bromide for 12 hours, preferably in a dark flask. The mixture is then decomposed with ammonium chloride solution, the ether solution is washed, dried and evaporated in vacuo. The residue is distributed between petroleum ether and 98 per cent. methanol, whereby the desired L18-di-[2,6,6trimethyl cyclohexene (1') yl] 3,7,12,16tetramethyl 8,11 dihydroxy octadecahexaene (2,4,6,l2, 14,16) -yne-(Q) goes over into the alcoholic aqueous phase. It is a highly viscous yellow oil with an absorption maximum at 280 ma (e=59000).

16 grams of the above'mentioned dihydroxy compound are dissolved in cc. of toluene and 300 mg. of p-toluene sulphonic acid in hot toluene are added at boiling temperature. The dehydration immediately takes place under vigorous boiling up while the solution turns red. The same is allowed to boil for l5 minutes, cooled down, Washed successively with bicarbonate and water, dried and evaporated carefully in vacuo at a maximum bath temperature of 50 C. The residue is dissolved in petroleum ether and chromatographed on a column of aluminium oxide (activity grade -II). The chromatogram is developed with increasing quantities of benzene, whereby a yellow first running as eluate and discoloured polymerization products in the upper part of the column are separated. At last, the orange coloured main zone is eluted with petroleum ether-alcohol. After the evaporation of the solvent, 2 grams of red oil remain behind which are taken up in carbon disulphide and alcohol is added. By evaporating a part of the carbon disulphide in vacuo, light red crystals are separated (900 mg.). Upon recrystallizing twice from benzene-methanol, metal-like shining red leaflets are obtained which melt in the evacuated tube at -156" C. Ultraviolet absorption maxima: 433, 458 my. (in hexane), 442, 4'10 my. (in benzene), 460, 490 ma (in carbon disulphide). -Colour reaction vwith antimonytrichloridein .ic'hloroformz At nrstgreen, then very slow- 1y llilue.

220 mg. ofV this .crystallisate (9,9.dehydroj,3- carotene) `are dissolved `in cc. .of Abenzene and, for puriiication purposes, left to Vstand for 12 hours with 200 mg. oi Pt-Si02 and 300 mg. of Rt-carbon. The mixture 'is iiItered, the filtrate is washed with 'l5 cc. o'fbenzene and the Aunited ltrates are Ahydrogenated by means of '609 mg. of Pd-CaCO-cata'lyst with the addition Lof 15 mg. of `quinoline. 1 rno'l of hydrogen ,isA taken up within minutes. The spectrum now .shows amain maximum at 450 m/randafsidemaximum at i340 ma. The catalyst is .sucked off, the ihydrogenation solution is .diluted with rhexane and washed With-dilute sulphuric acid .andwater In order to isomerize the product, the volume of the solution .is rst -brought to 600 cc.'by the addition of hexane and then allowed to stand 'for one hour in portions of 100 cc. with .'lOOfy each of iodine in 1 cc. of hexane. Thereupon, the mixture lis immediately chromatographed on a column o aluminium oxide (activity grade 4I--II). The orange zones of the trans--carotene which are .formed after .the .development with petroleum .ether-benzene (10:1.) .are .united .and eluted with petroleum etheiealcohol. After evaporation ofthe solvent in vacuo `.at low temperature .there remains .behind a crystalline .residue which is recrystallized from benzene-methanol. The violetredlealiets obtained melt in evacuated tubes at 179-180 C. and show no depression of melting point when mixed with .natural all-trans-- carotene (melting point 17.8 0.). Ultraviolet absorption maxima 452, 480 ma (in hexane). The cis-isomers obtained as .by-products yield, upon renewed isomerization with iodine, a further quantity of pure trans--carotene. The yield can furthermore be increased .by repeated chromatography of the mother liquors. Total yield of .pure .all-trans--carotene from dehydro- -carotene: 33 per cent.

EXAMPLE '3 3 parts by weight of 9,9"-deyhdro-carotene `of melting point 156 C. (prepared .according to Example '2) are '.hydrogenated., 4`under exclusion of light and at ythe 'lowest possible temperature, in benzene by .means of partially .poisoned palladium-calciumcarbonate-catalyst After l 4mol of hydrogen has been taken up, the catalyst .is ltered olf, Ythe 'benzene `solution is concentrated in vacuo and .crystallization is brought about by sprinkling with methanol. The crystals are sepai rated in .a centrifuge, washed with methanol `and further recrystallized twice from benzenemethanol in a few minutes. The '9,9'monocis lf2-carotene dried in high vacuo at 20 C. melts at 151 C. .Absorption maxima in hexane: 3.38 .ma (e=52000), .450 ma .(6:9255'00) and y-4'77 ima '(6:76'090).

l par't by weight .of .cis--.carotene in :10D-250 parts by volume of .hexane under carbonic acid .exposed to diiuse vda -light for 2 4 hours. The solution thereupon shows :the jpure absorption spectrum of all-trans- -carotenel In order to isolate the -carotene 'form-ed,the'hexane solution is adsorbed onto a column of aluminium oxide and the chromatogram is developed with cyclohexane-benzene. The orange coloured main zone is eluted with cyclohexane-alcohol. After evaporation of the solvent in vacuo, red crystals of melting point 178 C. are obtained from benzene-methanol; absorption maxima in hexane:

l4.52rmrand 480p. The yield, `calculated from cis- -carO'tene, is almost quantitative.

EXAMPLE 4 1 part by weight of 'cis--'carotena prepared according to Example 3 is heated, .under carbonic acid on a water-bath `in vthe dark, in 1D0-250 parts by volume of hexane for "2-'3 .hours at -*10 C. Thereupon, the solution .shows lsubstantially the absorption spectrum of the all-'trans-- carotene. For further working up, the vindications of Example 3 can .be followed.

8:4 grams Aof 1,18-di- [2,66-trimethylcyclo hexene `('1) -yll `3,"7,12,16 tetramethyl 8,11"- dihydroxy octadecahexaene (2,4,6,'l2;14,16)1 yne-(9) (prepared according to Example 1 vo1' 2) are dissolved in 5-0 cc. of vmethanol and, for purification purposes, allowed vto stand for 12 hours with 5 grams ofplatinized ycharcoal k(5 per cent.-). The coal is sucked off and washed vwith cc. lof methanol. After addition of 0.'5 gram of quinoline and 1.6 grains lof palladium charcoal (4"per eentp), the mixtureis partially hydrogenated until per cent. of 1 iinol of hydrogen have been taken up. After filtering Yoff the catalyst, 3 0'0 cc. of water vare :added and vextracted with ether. The .ether solution is washed 'with 1"Nsulphuric acid and water, dried and evaporated. The residue, a viscous yellow oil, is vdissolved in 50 cc. vof ytdluene and a hot `solution of 200 mg. of toluene sulp'honic acid in 30 cc. vof toluene is added at boiling heat under a nitrogen atmosphere. The solution immediately turns red. lIt is heated 'to ibo'iling for "I5 minutes, after cooling 'diluted Awith T00 cc. 'of .petroleum ether, washed successively with bicarbonate and water, dried Aand boiled down. The iedresidue is vtaken up in 50 cc. of petroleum yether and the solution is .shaken threetimes with 30 cc. each of y per cent. methanol. Thereupon, the petroleum ether solution 'is washed twice with water, dried .and Achromatographed on a column (3 x 40 cm.) of aluminium oxide (activity grade I-II). Already during "the development with petroleum ether, yellow Acoloured vcleavage products go into the filtrate. By petroleum ether with .an addition of 10 'per centi-benzene, further yquantitiesofiyellow by-products are eluted. An orange zone of an approximate 'breadth of A8 cm. can now be recognized in the middle vof the column underneath of which lighter coloured, broad, yellow zones 'are joined. In 'the 'upper lpart of the co1- fum'n, dirty coloured `resinous zones can be recognized. EThe column is now blown dry with the aid of nitrogen Aand divided. The vOrange zonefis eluted with petroleum ether-alcohol '(20:1l. 'The 'solutionshows the absorption maxima of -carotene which however are 'of low intensity. 'The alcohol is removed by washing with water and the dried petroleum ether solution 'is once vagain chromatographed on an aluminium -oxide column of V2 x 30 cm. (activity grade I-II). The lchromatogram is developed with increasing quantities of benzene until an orange zone distinctly {separates. This orange -zone is 'cut -out land eluted with-ether. The `solution shows the absorption maxima'of ,li-carotene. After evaporation of the fetherdn vacuo, the red lresid-ue is ltaken up in 2 cc. ofcarb'on disulphide and -8 ce. ef afledholare added. Then the mixture is concentrated cautiously until it starts to become cloudy. After standing overnight in an ice-box, red crystals separate. Gross yield: 20 ing. After two recrystallizations from benzene-methanol, the redviolet leaflets melt at 178179 C. maX.=452,480

(1,6) -yne-(B) in the ,compound selected from 11 The melting point of the mixall-trans--carotene shows no ma (in hexane). ture with natural depression.

EXAMPLE 6 Aand shaken out four times with 25 cc. each of 95 per cent. methanol. nish 2.7 grams of 1-methoxy2,6dimethyl5-hy- The methanol extracts furcyclohexenyl octadieneform of a viscous yellow oil. The determination according to Zerewitinoi shovvs 1 atom of active hydrogen.

droxy 8 trimethyl trimethylcyclohexeneyne (3) are dissolved in 6 cc. oi toluene and added dropwise to 1.2 grams of phosphorus oxychloride in 8 cc. of toluene and 4 cc. of pyridine. warmed to S30-95 C. for 45 minutes, poured onto ice after cooling and Washed with dilute sulphuric acid while adding ether. The ether layer is washed neutral, dried and evaporated. The residue is dissolved in petroleum other and chromatographed on aluminum oxide. By the aid of petroleum ether 0.8 gram of a yellow oil are eluted which, according to the Working method described in Example 1, are transformed into the 2,6-dimethyl- 8 [2',6',6'trimethylcyclohexene (1' yl] octadiene (5,7)yne (3) al- (1) (Ultraviolet absorption maximum oi the phenylsemicarbazone derivative at 275 ma and 359 mp) or into the dihydroxytetraenetriyne compound `:Vith 4() carbon atoms and hereafter into -carotene.

We claim:

1. A process which comprises subjecting a the group consisting of l-hydrocarbonoxy 2,6-dimethyl 8-l2,6,6'

.(l,3,5,7) and 1-hydrocarbonoxy-2,6dimethyl-x3- triene-(1,5,7)yne(3) to acid cleavage to produce a second compound respectively selected from the group consisting of 2,6- dimethyl-8- 1 .l2/,626 Atriene (2,4,6)

trimethylcyclohexene (1')-yl]octa -al-(l) and 2,6 dimethyl 8- [2,6',6'trimethylcyclohexene (1')-yl]- octadiene-(5,7) -yne-(3) -al-(l), condensing said second compound with acetylene to produce a third .compound respectively selected from the group -yne- (9) and 1,18-di- [2',6,6' -trimethylcyclohexkene-(1') -yll Vtreatment which comprises successive steps of methoxy 2,6 dimethyl 5- a The mixture is dehydration and hydrogenation to produce 9,10- rnono-cis L18-di-[2,6,6'trimethylcyclohex ene (1')-yl] -V 3,7,12,16 tetramethyl-octadecanonaene (1,3,5,7,9,11,13,15,17), and isomerizing the latter to produce all-trans--carotene.

Z. A process which comprises subjecting 1-hydrocarbonoxy 2,6 dimethyl-8[2',6',6' trimethylcyclohexene (1') yl] octatetraene- (1,3,5,7) to acid cleavage to produce 2,6-dimethyl 8 [2,6',6'-trmethylcyclohexene-(1')-yl] octatriene (2,4,6)al(1),,condensing the latter with acetylene; to produce 1,18-di-[2,6',6' trimethylcyclohexene- (1')-yl] 3,7,12,16tet ramethylY 8,11 dihydroxy f .octadecahexaene- (2,4,6,12,14,16) yne-(9), subjecting the latter to dehydration to produce 1,18-di-i2'6,6'tri methylcyclohexene (1')-y1] 3,7,l2,16tetra methyloctadecaoctaene (1,3,5,7,11,13,15,17) yne(9), rhydrogenating the latter to produce 9,10-mono-cis-1,18di [2!,6,6trirnethylcyclo hexene(1) yl]-3,7,12,16 tetramethyl-octadecanonaene (1,3,5,7,9,11,13,15,17), and isomerizing the latter to produce all-trans--carotene. A

3. A compound selectedfrom the group consisting of 2,6 dimethyl g 8-[2,6,6'trimethyl cyclohexene (1')-yl]-octatriene-(2,4,6)-al-(1) and 2,6-.dimethyl 8-[2,6,6 trimethylcyclohexene (1')yl]octadiene (5,7) -yne-(B) -al- 4. 2,6-dimethyl 8-[22626'A trimethylcyclohexene (1') -yll -octatriene-(2,4,6) -al-(l) 6. A compound selected from the group consisting of L18-di-[2',6,6trimethylcyclohexene (1') yl]3,7,12,16 tetramethyl 8,11 dihydroxy octadecahexaene (2,4,6,12,14,16)yne (9) and L18-di-[2',6,6'-trimethylcyclohexene- (1')-yll-3,7,12,16tetramethyl 8,11-dihydroxyoctadecatetraene (1315,17) triyne-(5,9,13).

7, 1,18-di [2,6',6' trimethylcyclohexene- (1') -yll V3,7,12,16-tetramethyl-,11-dihydroxyoctadecahexaene (2,4,6,12,14,16) -yne-(S).

9. 9,10-mono-cis1,18di[2',6,6 trimethylcyclohexene (1')-yl] -3,7,12,16 tetramethyl- .octadecanonaene- (.1,3,5,7,9,11,13,15,17)

HANS HERLOFF INHOFFEN. FERDINAND BOHLMANN.

References Cited inthe lepf this patent UNITED STATES PATENTS Number Name Date 2,369,159 Milas Feb. 15, 1945 2,451,735 Isler Oct. 19, 1948 2,451,736 Isler Oct. 19, 1948 `2,451,737 Islerp-; Oct. 19, 1948 2,451,738 Isler Oct. 19, 1948 2,529,498V Isleri.;v Nov; 14, 1950 OTHER REFERENCES Inhoffen et al., AChem. Abstracts, vol. 44, col. 8890,-1 page only (1950). 

4. 2,6-DIMETHYL - 8-(2'',6'',6'' - TRIMETHYLCYCLOHEXENE - (1'') - YL)-OCTATRIENE- (2,4,6) - AL-(1).
 5. 28L - DIMETHYL-8-(2'',6'',6'' - TRIMETHYLCYCLOHEXENE - (1'')-YL)-OCTADIENE - (K,7)-YNE-(3) -AL(1). 